BACKGROUND: Arrhythmogenic right ventricular cardiomyopathy (ARVC) has been linked to mutations in desmosomal proteins, including plakophilin-2 (PKP2). Little is known about the changes in cellular function and structure that follow expression of ARVC-relevant PKP2 mutations. OBJECTIVE: The purpose of this study was to investigate the function and distribution of an ARVC-relevant PKP2 mutant where arginine at position 79 was replaced by a stop codon (R79x). METHODS: Results were compared with those obtained with mutation 179fs (frameshift at position 179). Mutant constructs were introduced by adenoviral infection into neonatal rat ventricular myocytes in culture. RESULTS: Both mutant proteins failed to preferentially localize to sites of cell-cell apposition. Their expression did not disrupt localization of endogenous PKP2, connexin-43 (Cx43), or desmoplakin (DP). However, we observed reduced abundance of Cx43 after R79x expression. Early truncation of PKP2 at position 79 also prevented its physical interaction with both DP and Cx43. Finally, R79x expression correlated with loss of expression of HSP90, a protein relevant to cardiomyocyte apoptosis. CONCLUSION: These results provide the first observations of the cellular/molecular phenotype consequent to these PKP2 mutations and give insight into the possible cellular substrates that lead to ARVC.
BACKGROUND:Arrhythmogenic right ventricular cardiomyopathy (ARVC) has been linked to mutations in desmosomal proteins, including plakophilin-2 (PKP2). Little is known about the changes in cellular function and structure that follow expression of ARVC-relevant PKP2 mutations. OBJECTIVE: The purpose of this study was to investigate the function and distribution of an ARVC-relevant PKP2 mutant where arginine at position 79 was replaced by a stop codon (R79x). METHODS: Results were compared with those obtained with mutation 179fs (frameshift at position 179). Mutant constructs were introduced by adenoviral infection into neonatal rat ventricular myocytes in culture. RESULTS: Both mutant proteins failed to preferentially localize to sites of cell-cell apposition. Their expression did not disrupt localization of endogenous PKP2, connexin-43 (Cx43), or desmoplakin (DP). However, we observed reduced abundance of Cx43 after R79x expression. Early truncation of PKP2 at position 79 also prevented its physical interaction with both DP and Cx43. Finally, R79x expression correlated with loss of expression of HSP90, a protein relevant to cardiomyocyte apoptosis. CONCLUSION: These results provide the first observations of the cellular/molecular phenotype consequent to these PKP2 mutations and give insight into the possible cellular substrates that lead to ARVC.
Authors: Brenda Gerull; Arnd Heuser; Thomas Wichter; Matthias Paul; Craig T Basson; Deborah A McDermott; Bruce B Lerman; Steve M Markowitz; Patrick T Ellinor; Calum A MacRae; Stefan Peters; Katja S Grossmann; Jörg Drenckhahn; Beate Michely; Sabine Sasse-Klaassen; Walter Birchmeier; Rainer Dietz; Günter Breithardt; Eric Schulze-Bahr; Ludwig Thierfelder Journal: Nat Genet Date: 2004-10-17 Impact factor: 38.330
Authors: Starr R Kaplan; Joseph J Gard; Luis Carvajal-Huerta; Juan C Ruiz-Cabezas; Gaetano Thiene; Jeffrey E Saffitz Journal: Cardiovasc Pathol Date: 2004 Jan-Feb Impact factor: 2.185
Authors: Sarah B Geisler; Kathleen J Green; Lori L Isom; Sasha Meshinchi; Jeffrey R Martens; Mario Delmar; Mark W Russell Journal: J Biomed Biotechnol Date: 2010-05-12